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The Impact of a Fungus-Feeding Nematode (Aphelenchoides Sp.) on Decomposition of Trembling Aspen Wood by Various Wood-Decay Fungi

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The Impact of a Fungus-Feeding Nematode (Aphelenchoides Sp.) on Decomposition of Trembling Aspen Wood by Various Wood-Decay Fungi Lakehead University Knowledge Commons,http://knowledgecommons.lakeheadu.ca Electronic Theses and Dissertations Undergraduate theses 2018 The impact of a fungus-feeding nematode (Aphelenchoides sp.) on decomposition of trembling aspen wood by various wood-decay fungi Reale, Julia http://knowledgecommons.lakeheadu.ca/handle/2453/4498 Downloaded from Lakehead University, KnowledgeCommons THE IMPACT OF A FUNGUS-FEEDING NEMATODE (APHELENCHOIDES SP.) ON DECOMPOSITION OF TREMBLING ASPEN WOOD BY VARIOUS WOOD- DECAY FUNGI by Julia Reale FACULTY OF NATURAL RESOURCES MANAGEMENT LAKEHEAD UNIVERSITY THUNDER BAY, ONTARIO May 2018 THE IMPACT OF A FUNGUS-FEEDING NEMATODE (APHELENCHOIDES SP.) ON DECOMPOSITION OF TREMBLING ASPEN WOOD BY VARIOUS WOOD- DECAY FUNGI by Julia Reale An Undergraduate Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Honours Bachelor of Science in Forestry Faculty of Natural Resources Management Lakehead University May 2018 ____________________________ ____________________________ Major Advisor Second Reader i LIBRARY RIGHTS STATEMENT In presenting this thesis in partial fulfillment of the requirements for the HBScF degree at Lakehead University in Thunder Bay, I agree that the University will make it freely available for inspection. This thesis is made available by my authority solely for the purpose of private study and research and may not be copied or reproduced in whole or in part (except as permitted by the Copyright Laws) without my written authority. Signature: _______________________________ Date: ____________________________________ ii A CAUTION TO THE READER This HBScF thesis has been through a semi-formal process of review and comment by at least two faculty members. It is made available for loan by the Faculty of Natural Resources Management for the purpose of advancing the practice of professional and scientific forestry. The reader should be aware that opinions and conclusions expressed in this document are those of the student and do not necessarily reflect the opinions of the thesis supervisor, the Faculty of Natural Resources Management or Lakehead University. iii ABSTRACT Reale, J. 2018. The impact of a fungus-feeding nematode (Aphelenchoides sp.) on decomposition of trembling aspen wood by various wood-decay fungi. 56 + viii Pp. Keywords: Aphelenchoides sp., Bjerkandera adusta, Cerrena unicolor, Climacodon septentrionale, fungi, Ganoderma applanatum, grazing, Hohenbuehelia grisea, mycophagy, nematodes, nematophagous, Sphaerobolus stellatus, Trametes pubescens, wood-decay. Grazing by fungus feeding invertebrates on fungal mycelium can potentially impact many important ecological processes such as the formation of mycorrhizas and the decomposition of wood and litter. A study was initiated to examine the impact of a fungus feeding nematode (Aphelenchoides sp.) on the decomposition of trembling aspen (Populus tremuloides) wood blocks under aseptic growing conditions by seven different decay fungi. These fungi were Bjerkandera adusta, Cerrena unicolor, Climacodon septentrionale, Ganoderma applanatum, Hohenbuehelia grisea, Sphaerobolus stellatus, and Trametes pubescens. Results based on dry weight measurements of wood blocks before and after inoculation and in the presence or absence of nematodes revealed that four of the seven fungi exhibited lower rates of decay in the presence of nematodes. These fungi were T. pubescens, G. applanatum, C. septentrionale and S. stellatus. The other three fungi had slight increases in decomposition of wood blocks in the presence of nematodes. It is suggested that B. adusta and C. unicolor may have responded to grazing by producing enhanced mycelial growth and thus enhanced enzymatic activity. Hohenbuehelia grisea is a known nematophagous fungi, capturing and consuming nematodes as a supplementary source of nitrogen, thus accounting for enhanced decomposition in the presence of nematodes. iv CONTENTS ABSTRACT III LIST OF TABLES VI LIST OF FIGURES VII ACKNOWLEDGEMENTS VIII INTRODUCTION 1 Wood Decay 1 Types of Wood Decay 2 White Rot 2 Brown Rot 3 Soft Rot 4 What Fungi Cause Decay? 5 Ecological Role of Wood Decay Fungi 6 Obstacles Faced 7 Objective 12 Hypothesis 12 MATERIALS AND METHODS 13 Wood block preparation 13 Flask preparation 13 Experimental Design 14 Inoculation of flasks with wood-decay fungi 16 Inoculation of Aphelenchoides sp. 17 Addition of extra water 19 Harvesting wood blocks 19 Statistical analysis 20 RESULTS 21 DISCUSSION 28 Trametes pubescens 31 Bjerkandera adusta 31 Ganoderma applanatum 32 Cerrena unicolor 33 Climacodon septentrionale 34 Hohenbuehelia grisea 35 Sphaerobolus stellatus 35 v CONCLUSION 37 LITERATURE CITED 39 APPENDIX I 47 APPENDIX II 48 APPENDIX III 51 APPENDIX IV 55 APPENDIX V 56 vi LIST OF TABLES Table Page 1. Experimental Design 15 2. Summary table showing average dry weights before and after inoculation 23 and their average percent differences (percent decay %) 3. Results from univariate analysis (one way ANOVA) 23 4. Treatments with their respective average percent decay (%) sorted from 25 smallest to largest, and LSD group vii LIST OF FIGURES Figure Page 1. Photo taken by Dr. Hutchison of Aphelenchoides sp. feeding on hyphae 11 2. Dried wood blocks being weighed 13 3. Flasks prepared with vermiculite-peat mixture and malt extract broth 14 before being autoclaved 4. Experimental design showing 10 flasks for each fungus containing no 16 nematodes, and other 10 containing nematodes 5. Author inoculating wood-decay fungi on wood blocks (left) and example 17 of Cerrena unicolor agar plugs from petri dish (right) 6. Author using Eppendorf micropipette to inoculate 200 nematodes/mL 18 of Aphelenchoides sp. (top) and sterilize flask using Bunsen burner (bottom) 7. 150 Erlenmeyer flasks in incubator for incubation at 20°C 19 8. Mycelium present in Erlenmeyer flasks before harvesting and drying 20 wood blocks for statistical analysis 9. Figure showing the percent decay (%) corresponding to each treatment, 26 with the LSD grouping labelled 10. Average percent decay (%) for each treatment of wood-decay fungi, 27 with the presence of nematodes shown in orange 11. Average percent decay (%) for each treatment of wood-decay fungi, 27 with the presence of nematodes shown in orange, versus no nematodes shown in blue viii ACKNOWLEDGEMENTS I wish to thank and express sincere gratitude to my thesis advisor Dr. Leonard Hutchison for his guidance, knowledge, enthusiasm, patience and support in helping me complete this thesis. I would also like to thank my second reader, Dr. Leni Meyer in helping me with statistical analysis. I am also grateful for my friends and family who have supported me along the way. 1 INTRODUCTION WOOD DECAY Wood is a very important substance in our world and is of considerable anatomical and chemical complexity; supporting rich communities of fungal species in a variety of microbial niches (Dix and Webster 1995). A key component in the healthy life cycle of a forest is the presence of wood-inhabiting fungi which are extremely important in the process of nutrient and carbon cycling in temperate forests. Wood rotting fungi are, by definition, those which can bring about significant weight loss and structural change in woody tissues (Dix and Webster 1995). Wood is composed of mostly cellulose, which is a polymer of sugars, and lignin which is a complex heterogeneous polymer that is made up of several phenol-containing compounds. Trees differ in their wood structure, as does the enzymatic potential of the fungi for decomposition. The degree of decomposition on the substrate and the method used to do so will depend on the ability to degrade different cell types and cell-wall constituents (Schwarze et al. 2000). Coarse woody debris makes a large contribution to immobilized nutrients such as Phosphorus (P) and Nitrogen (N) that can be readily utilized by wood-decay fungi (Stenlid et al. 2008). Fungi which decay wood are some of the few organisms that can utilize these nutrients by secreting an array of enzymes with the unique ability to disassemble the complex molecules that comprise wood (cellulose, lignin and hemicellulose) and recycle it back into the ecosystem (Boddy and Watkinson 1995). 2 TYPES OF WOOD DECAY However, not all fungi secrete the same array of enzymes to disassemble wood, and thus not all fungi attack wood in the same way. They may be distinguished by differences in colour, solubility, strength, dimensional stability, pulping properties and the chemical composition of the decayed wood (Cowling 1961). As a result, one might expect to see one of the following three types of rot: white rot, brown rot and soft rot depending on the enzymatic arsenal of the fungus and the host that it is attacking (Dix and Webster 1995). In some cases, wood-decay fungi may use several different methods of attack on the same tree (Dix and Webster 1995). White Rot White rot is the most common type of decay and normally leaves a bleached appearance due to the oxidation of lignin in wood and may occur uniformly, leaving the wood spongy or stringy. White rot may also appear as a selective decay or a pocket rot (Goodell et al. 2008). White rot fungi possess both cellulolytic and lignin degrading enzymes, therefore having the potential to degrade the entirety of the wood structure under the correct environmental conditions (Goodell et al. 2008). White rot fungi are the most efficient lignin degraders in nature, but erode the cellulose and hemicellulose components in wood as well (Dix and Webster 1995). White rot can also
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